LOCUS       sequence-227218-        7436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RPCC7116RT-Cas1-Cas2 under pT7lac promoter, encodes 
            RPCC7116CRISPR array 2 RC, compatible with SENECA acquisition 
            readout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7436)
  AUTHORS   Schmidt F, Cherepkova MY, Platt RJ
  TITLE     Transcriptional recording by CRISPR spacer acquisition from RNA.
  JOURNAL   Nature. 2018 Oct;562(7727):380-385. doi: 10.1038/s41586-018-0569-1. 
            Epub 2018 Oct 3.
  PUBMED    30283135
REFERENCE   2  (bases 1 to 7436)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-018-0569-1"; journalName: "Nature"; date: "2018-10";
            volume: "562"; issue: "7727"; pages: "380-385"
COMMENT     Sequence Label: 
            pFS_0388_pET30-RT-CRISPR_Rivularia_sp._PCC_7116_ARRAY_2_RC
FEATURES             Location/Qualifiers
     source          1..7436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(359..1174)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(359..1174)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1082..1101
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1166..1185
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1296..1884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1785..1804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2038..2055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2070..2209
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2135..2154)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2295..2317
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2311..2502)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3311..4393)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4355..4374
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4394..4471)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4677..4696
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4784..4803
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4784..4802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4803..4827
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             4842..4864
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(7349..7367)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7363..7410
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ccaggttaag ttaattacag cttgattttt catccaattt ttatattaaa agcctaccgc
       61 ttaaattggt aggctttttt attgcaagtc attgtaactc tttactgttt ccaactaatc
      121 cgaattaacc caatcggtgg gggtcccgcg gccgctattc ttttgattta taagggattt
      181 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      241 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
      301 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
      361 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
      421 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
      481 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
      541 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
      601 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
      661 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
      721 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
      781 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
      841 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
      901 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
      961 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1021 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     1081 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     1141 tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     1201 aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga
     1261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1441 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggtc
     1921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2041 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     2101 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2161 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     2221 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2281 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2341 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     2401 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     2461 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     2521 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     2581 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     2641 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     2701 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     2761 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagatcat
     2821 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     2881 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     2941 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
     3001 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3061 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     3121 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagatagt
     3181 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3241 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3421 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3481 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3541 gcgggatata acatgagcta tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3601 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3661 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3721 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3781 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3841 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccat
     3901 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3961 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4021 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4081 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4141 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4201 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4261 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4321 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4381 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4441 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4501 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4561 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     4621 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     4681 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     4741 cgtccggcgt agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata
     4801 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     4861 gagatataca tatgggatac gagaacatca gataccttca caagccgtac ttcgcagaaa
     4921 cgattactga gaagccacca cagactgagg aagatgactc accattccag aagaacatct
     4981 ggaacgctga gatggctgca atctatctta tcgagcaggg tacaaacatc tacaaagact
     5041 accagcgttt cataatacac gttagtgaga agccaaaact tgaggtccct atcagggacg
     5101 tacaacagat catcgtcttt ggaaatatcc agctaagcac gcctgttatc caggcctgcc
     5161 tgaaggagca gatcccagtt gttttcctgt ctcagaccgg aacataccat ggccatctgt
     5221 ggtcggagaa gtcaatccac cttgataacc agttggttca ggctgagcga aggaacgacg
     5281 acttattcag attctcagtt agccgagcag tggttctagg gaagcttctg aactctaagc
     5341 aactactgat gagattcaac agaaggagaa agattggtaa ggtcgaagag gcaatctacg
     5401 gtatcaacca ggacatagac gcattgaatt acgtggacaa cctggacacg ctgaggggat
     5461 acgagggtat cgcagccgcc cgttacttcc ccgccttcgg taacttgata acaaacccaa
     5521 agttcagctt cagtcaaagg ttcaggcagc caccaactga cgaaattaac tctctactgt
     5581 ccttcggtta cacattgtta ttcaacaacg ttttatcctt catcatcact gagggcttgt
     5641 cgccgtacat aggacacttc cattacggtg acaagcagaa aacttacctc gcattcgacc
     5701 ttatggaaga gttcagatcg cctatcgtag actctcttgt attgaagata attaacaaga
     5761 gcctattcaa gccccaagac tttgacgttg tcgcctctac cggcggtgtt tatttgtctc
     5821 agaccagccg tagggtgttc ctgaagcaat tcgagaacag aatgaacgag gaaatctccc
     5881 acccggacct gatctcccag gttacttacc gtcatgctat ccagttgcag gtcagacgat
     5941 acaaaaggtg cctgctgagt gacaacatct acgagtcttt cctaagggct gactaattaa
     6001 ctttaagaag gagatataca tatgatgata tcatctcagt tcatagacat aaacaacttc
     6061 cagagagcat gggagaaagt cgcagacaag agagggtgtg ctggagtaga cggagagaca
     6121 atctcctcat tcgccagtaa ccagactgtg aacgtctacc aactcatgaa tagtgtggca
     6181 gacggatctt accagccgtt cccatgtaaa caagtcatca tcccaaagag gaatgggtcc
     6241 cagagggaac taaagatacc gaccatcagg gataggatag tgcagcaggc tctcctaaac
     6301 gtcatctcac ctcttatgga agaaaagttc tctccagtct ctttcgcata caggccaaac
     6361 ttgagctaca ttaacgccgt ggagaagatc gcagattgga gagatatggg gtacgtttgg
     6421 gtcctggacg ccgacattgt gaaattcttc gacaacatcg atcaccacag acttttgcag
     6481 caagtgcgac tacacatcga tcaccctggc atcctgtgcc tgataaaggc atggatctct
     6541 gtcggcgtcg agacacgaga gggtttgatt cttccacaga aaggaatccc acagggcgct
     6601 gtcatctcac ctatcttagc taacatctac ctgcacgagt tcgacgagat aatctctgca
     6661 tctgacctag agatagtaag atacgccgac gacttcttag tactatccac cagccaggag
     6721 agaatagcca tagccaagtc ccaggttata gacttgctcg actcattggg tctagagatt
     6781 aacacagaca agacacagat cacgtctttc gagagaggat tcagattcct tggacacggg
     6841 tttctgtccg acgcaatatt tccagtggac accaacaagg ctaaactgaa aagtggaatc
     6901 gagacaaaca gagaaaagac taggacccga aagacgagca agaagaagct ctaccataat
     6961 ccttacagaa ataagaaggt cgtatgacgt acgttaactt taagaaggag aacttaagat
     7021 gcttgtggtg gtggtttatg acattcccaa tgacaagagg cgaacaaaac tgtctaactt
     7081 cttggaaggg tacggacagc gtgtgcagtt ctcagttttc gagtgcttct taaacctgga
     7141 cgaaatgaga cagctattcg agaaaagcaa gaagattgta aagccgtctg aggataacgt
     7201 gagattctac tggatatcag aggacgcagt gtctcgtgtg cttaccatag gctctgagca
     7261 ccccaatgcc ccacctaatt attacgtcat ctaactcgag atccggctgc taacaaagcc
     7321 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     7381 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggata
//
